SENIOR HIGH SCHOOL DEPARTMENT

LEMU NATIONAL HIGH SCHOOL

“ANTIBACTERIAL ACTIVITY OF A NATURAL ACLEO

LIQUIGEL AGAINST STAPHYLOCOCCUS AUREUS”

A Research Paper

Presented to

The Faculty of the Senior High School Department

LEMU NATIONAL HIGH SCHOOL
Lemu Sur, Enrile, Cagayan

In Partial Fulfillment of the Requirements

of the Subject Inquiries, Investigations and
Immersion

STEM – GRADE 12

S.Y. 2020-2021

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PREPARED BY:

RITCHIEDON LABBAO TURARAY

GERALD PALATTAO ANDRIN
ALBERTSON PERPINIAN BINASOY
KARYLLE BINARAO LONZANEDA
LOVELY JOY GONZALES GUINUCUD
RYAN DATUL MARTINEZ
KAYE ANNE LOLO PALATTAO
NEIL JOHN GUZMAN TUPPAL
MEYNARD TELAN CAGURANGAN
MAGDALYN BAQUIRAN MABBORANG
PETER JOHN LOTILLA GUMABAY
ELJHAY BUQUEL MOJICA
QUINEE JOY BASSIG ACORDA
AISYD RIANNE MACARILAY TAMON
NATHANIEL SEGUI CASAUAY
JIMMY BACCAY MABBORANG
CLARK SALVADOR BUNAGAN
JEHARD AGUB SIBAL
NEAL CHESTER SINGAYAN MABBORANG
ERICA LOVE DECENA BACUD
ROVINCE GUINUCUD MADDAWIN

EVA PLOPINIO SALES

INQUIRIES, INVESTIGATION AND IMMERSION TEACHER

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CHAPTER I
INTRODUCTION

One of the greatest problems that we are facing in our

community, country and also the whole World is the spreading
of Corona Virus or the commonly called COVID-19. Coronavirus
disease (COVID-19) is an infectious disease caused by a
newly discovered coronavirus. Most people infected with the
COVID-19 virus will experience mild to moderate respiratory
illness and recover without requiring special treatment.
Older people, and those with underlying medical problems
like cardiovascular disease, diabetes, chronic respiratory
disease, and cancer are more likely to develop serious
illness. The best way to prevent and slow down transmission
is to be well informed about COVID-19 virus, the disease it
causes and how it spreads. Protect yourself and others from
infection by washing your hands often with soap and water or
clean them with an alcohol-based sanitizer. This kills
viruses on your hands, practice social distancing because
you can have and spread the virus without knowing it, you
should stay home as much as possible. If you have to go out,
stay at least 6 feet away from others. And most important is
using of facemask, masks should be used as part of a
comprehensive strategy of measures to suppress transmission
and save lives, the use of mask alone is not sufficient to
provide an adequate level of protection against COVID-19.
The COVID-19 virus spreads primarily through droplets of
saliva or discharge from the nose when an infected person
coughs or sneezes, so it’s important that you also practice
respiratory etiquette (for example, by coughing into a
flexed elbow). Make wearing a mask a normal part of being
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around other people. The appropriate use, storage and

cleaning or disposal of masks are essential to make them as
effective as possible.
One of the recommendations of World Health
Organization (WHO)is to maintain hand hygiene in order to
decrease the risk of gastoenteric and respiratory
infections. Hence, this research is very important nowadays
to combat COVID-19 pandemic.
The researchers will seek the antibacterial activity
of ACLEO (ALOE VERA, CALAMANSI ZEST, AND LEMON ESSENTIAL
OIL) Liquigel against Staphylococcus Aureus. If this study
will be effective, the researchers will give help to those
who are deprived financially to prevent gastoenteric and
respiratory infections.

OBJECTIVE OF THE STUDY

In general, this study will assess “Antibacterial

activity of a natural ACLEO Liquigel against Staphylococcus
Aureus”

Specifically, this study aimed to:

1. Determine the zone inhibition of Aloe Vera (Aloe
Barbadensis Miller) with Calamansi (Critrofortunella
Microcarpa) with Lemon (Citrus Limon L.) Essential Oil.
a. 25%
b. 50%
c. 75%
2. Determine the zone inhibition of Aloe Vera (Aloe
Barbadensis Miller) with Lemon (Citrus Limon L.) Essential
Oil.

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a. 25%
b. 50%
c. 75%
3. Determine the presence of Yeast Escherichia Coli and
total Yeast count in the ACLEO Liquigel.
4. Determine the significant difference of Aloe Vera with
Calamansi with Lemon essential oil in terms of:
-Zone inhibition compared to positive control
5. Determine the significance difference of Aloe Vera with
Lemon essential oil in terms of:
-Zone inhibition compared to positive control

HYPOTHESIS

1. There is no significant difference in the mean zone of

inhibition of Aloe Vera (Aloe Barbadensis Miller) with
Calamansi (Critrofortunella Microcarpa) with Essential Oil.

2. There is no significant difference in the mean zone of

inhibition of Aloe Vera (Aloe Barbadensis Miller) with
Calamansi (Critrofortunella Microcarpa) with Lemon (Citrus
Limon L.) Essential Oil.

SIGNIFICANCE OF THE STUDY

This study will evaluate the “Antibacterial Activity of

a Natural ACLEO Liquigel Against Staphylococcus Aureus”
which is one among the pathogens that cause skin infection.
This study will also eradicate unwanted side effect caused
by antibacterial agents.
This study will be significant to the following:
To the Community people:
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This study will explore to lead a hand to indigent

people who cannot afford liquigel for them to have an
alternative antibacterial out of aloe vera, calamansi, and
lemon essential oil.
To the Researchers:
This study will discover alternative antibacterial and
to help combat the antibacterial resistant.
To the Future Researchers:
This study will serve as the guiding material for
future researchers. They could use, revise or add something
better in our procedure in preparing and making liquid gel.
This will also serve as a basis for the future researchers
who want to conduct the same study.
Thus, this study will be very useful and beneficial.

SCOPE AND DELIMITATION

This study is limited to the evaluation of

“Antibacterial Activity of a Natural ACLEO Liquigel Against
Staphylococcus Aureus”.

DEFINITION OF TERMS

Aloe vera
 Conceptual: Scientific name of Aloe Barbadensis, Aloe
vera is a popular medicinal plant that people have used
for thousands of years.
 Operational: It is a thick, short-stemmed plant that
stores water in its leaves.

Gel

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 Operational: colloidal dispersion which may be jelly-like

or not.
Calamansi
 Conceptual: scientific name Critrofortunella microcarpa,
this fruit is a hybrid of a member of the citrus genus,
that is to say, orange and kumquat, and is widely
cultivated in Southeast Asia, particularly Philippines.
 Operational: a ball-shaped citrus fruit which is green
when raw and yellow when it is riped; can be found within
the locality.
Lemon
 Conceptual: (Citrus ×limon), small tree or spreading bush
of the rue family (Rutaceae) and its edible fruit.
 Operational: A yellow, oval citrus fruit with thick skin
and fragrant, acidic juice.
Antibacterial Activity
 Conceptual: ability to inhibit or kill the growth of
bacteria that causes diseases to human.
Zone of Inhibition Test
 Conceptual: (also known as the Kirby-Bauer Test,
Antimicrobial Susceptibility Test, Disk Diffusion Test,
or Agar Diffusion Test) is a quick way to assess the
antimicrobial activity of a material or solution in
relation to a target microorganism.
ANOVA
 Conceptual: Analysis of Variance is a statistical method
that separates observed variance data into different
components to use for additional tests.
CHAPTER II

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REVIEW OF RELATED LITERATURE

This section contains the review of related literature

and studies. This includes the uses of aloe vera and
calamansi, properties of aloe vera and calamansi, benefits
of aloe vera and calamansi

Properties of Aloe Vera

Aloe vera properties make them very versatile. When
ingested, they are incredibly soothing, loaded with
antioxidants, and offer countless other health benefits. Of
course, the various species of aloe will be composed of
different compounds and properties. Even the inner and outer
parts of aloe leaves contain different properties. The
biggest between most of them is typically the amount and
density of minerals, vitamins and nutrients (Dr.
Edward,2016). In addition, aloe vera properties includes
Anti- Pruritic Properties and Healing Properties.

Anti-Pruritic Properties
Anti-pruritic properties are those that can alleviate
itching and dryness. Pruritus ailments include vexing skin
and scalp issues, like psoriasis and eczema. Psoriasis is an
auto-immune disease characterized by the excessive
production of skin cells. Recall, Aloe Vera’s enzymatic
properties, enabling Aloe to destroy excessive dead skin
cells, while conditioning the skin and scalp.
Since excess skin cell production is the heart of Psoriasis’
existence, you could see why Aloe is able help eliminate
this issue. Aloe can also reduce redness, scaling, itching
and inflammation of scalp psoriasis, which directly benefits
the hair, as well (Anonymous, 2012).

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Healing Properties
Aloe is an excellent treatment for skin conditions such
as burns and eczema. It is often reported that burns can be
healed remarkably quickly and the pain reduced very quickly
with topical application of Aloe Vera to the burn area. As
well as applying topically, Aloe can also be taken
internally so it just as useful for internal epithelial
tissues as it is for the skin. For example, mouth and
stomach ulcers, nasal, and sinuses, bowels lungs and genital
tract (Anonymous,2012).

Uses of Aloe Vera

Aloe Vera is one of the most powerful and well-known
medical plants. It has been used for more than 5,000 years,
and traces of it have been found in numerous civilizations.
Today, it has a strong global reputation due to widespread
usage in cosmetics, particularly to treat burns and sunburn,
to aid wound healing, and fight against cell aging. It also
helps in reinforcing the immune system and improve blood
circulation. In addition, aloe vera gel can relieve sunburn
and help heal wounds. But this plant can also use for much
more than sunburn and household reliefs (Anonymous,n.d.).

The following are the amazing uses of aloe vera:

Heartburn relief – A 2010 review suggested that consuming 1
to 3 ounces of aloe gel at mealtime can reduce the severity
of GERD (gastoroesophageal reflux disease). It can also ease
other digestion-related problems. Aloe vera’s low toxicity
makes it a safe and gentle remedy of heartburn.

Keeping produce fresh – Aloe gel can help fruits and

vegetables stay fresh, and eliminate the need for dangerous
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chemicals that extend the shelf life of produce. Aloe gel

coating successfully blocked the growth of many types of
harmful bacteria on the fruits and vegetable.

An alternative to Mouthwash - Aloe vera extract is a safe

and effective alternative to chemical-based mouthwashes.
Researchers found out that aloe’s natural ingredients, which
include a healthy dose of vitamin C, can block plaque. It
can also provide relief if you have bleeding or swollen
gums.

Lowering blood sugar – Aloe vera may have a future diabetes

treatment according to Phytotherapy research that used pulp
extract. People with diabetes who take glucose-lowering and
medications, should use caution when consuming aloe vera.
The juice along with diabetes medications could possibly
lower your glucose count to dangerous level.

A natural laxative – Aloe vera is considered a natural

laxative. A handful of studies have looked into the benefits
of the succulent to aid digestion.

Skin care – Aloe vera is helpful to keep your skin clear and
hydrated. Its water dense leaves, combined with special
plant compounds called complex carbohydrates, make it an
effective face moisturizer and pain reliever.

Benefits of Aloe Vera

9 Proven Aloe Vera Benefits
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Here are nine proven aloe vera benefits from Dr. Josh Axe,
aloe vera can soothes rashes and skin irritations. In
addition, it treats burns, heals cold sores, and moisturizes
hair and scalp. Treats constipation, helps with digestion
boost the immune system and provides antioxidants. Lastly,
reduces inflammation and treats diabetes.

Other Benefits of Aloe Vera

Aloe vera is high vitamins and minerals. Aloe vera is
also high in amino acids and fatty acids. Aloe vera is an
adaptogen, helps with detoxification and alkalizes the body.
Cardiovascular health, great for skin disinfectant, anti-
biotic, anti- microbial, germicidal, anti-bacterial, anti-
septic, anti- fungal and anti- viral. Lastly, weight loss- a
secondary effect (Dawn,2012).

Properties of Calamansi
Aromatic, antiseptic, antiphlogistic, carminative,
deodorant, refrigerant. Studies have suggested
antimicrobial, antianxiety, antidepressant,
hepatoprotective, expectorant and antioxidants properties
(Straut Jr.,2016).

Angiogenesis or the development of blood vessels and

oxidation or the radical transformation of cellular
substances are two key processes that contribute to cancer-
to-cancer progression. Hence, the study that evaluated the
antiangiogenic and antioxidant properties of different
concentrations of calamansi (citrusmicrocarpa) enthanolic
peel extract can lower the number of blood vessels that can
trigger cancer progression and more powerful antioxidants
property than the ascorbic acid (Anonymous,2016).

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Uses of Calamansi
Calamansi is one of the tiniest and cheapest fruit that
you will see in the market. It is a small fruit but it does
not reflect the many benefits it can give your body. This
fruit helps in enhancing beauty, helps promote your immune
system and help you lose weight (Yayang,2014).

The following are the other uses of calamansi, calamansi can

be used for hair growth. Before taking bath, rub calamansi
in your scalp and after 15 minutes you can already rinse it.
Do it every day to help your hair grow. If you have
dandruff, you can use calamansi to reduce it. Just rub it to
your scalp to reduce itching. It can be also used as a
deodorant. After taking a bath, rub the slice calamansi in
your armpit and let it dry. It will also help lighten your
armpit because sometimes deodorants tend to darken our
armpits. It helps whiten your skin. If you are dizzy or
nauseous, you can smell the peel of calamansi just squeeze
it, just be careful not to squeeze it near your eyes. It can
also remove stains in your clothes. Just dab some calamansi
drops on stained area. It also helps in whitening white
clothes and also it helps removed acne (Yayang,2014).

Benefits of Calamansi
9 Benefits We Can Get from Calamansi
The benefits of calamansi, useful for food recipes,
eliminate bad smell on fish, skin lightening and take away
stains on the clothes. Moreover, it washes body dirt, can be
used as deodorizer, eliminate itching and promotes hair
growth. Helps cough and good source of vitamin C
(Anonymous,2015).

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Amazing benefits of Calamansi

Calamansi improves your skin condition and it lowers
weight. Eliminates body odor, it works as a condiment and it
protects your teeth. It also protects you from flu and colds
and it clears dandruff (Anonymous,2014).

Other Health Benefits of Calamansi

Calamansi helps strengthen our immune system. It has
vitamin C that helps our bones and teeth healthy. It helps
quicken the body’s ability to repair wounds. Calamansi juice
serve as a liver clean. It helps eliminate toxins in our
body that results in faster weight loss. It helps reduce
burning sensation. It helps increase perspiration that help
lessen the uncomfortable effects if you have colds, flu and
fever. It helps control high blood pressure. A mixture of
calamansi and water can help a person with respiratory
problem (Yayang,2014)

Lemon
The main objective of the study entitled “Study of the
Antimicrobial Activity of Lemon (Citrus lemon L.) Peel
Extract” conducted by Maruti J. Dhanavade, Chidambre B.
Jakulte, Kailash D. Sonawane, Jai S. Ghosh, is extraction,
identification of antimicrobial compounds and demonstration
of antimicrobial activity of lemon (Citrus lemon L.) peel
against bacteria. As microorganism are becoming resistant to
present day antibiotics, our study focuses on antimicrobial
activity and future prophylactic potential of the lemon
peel. Biologically active compounds present in the medicinal
plants have always been of great interest to scientist. The
peel of citrus fruits is a rich source of flavanones and
many polymethoxylated flavones, which are very rare in other
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plants. These compounds, not only play an important

physiological and ecological role, but are also of
commercial interest because of their multitude of
applications in the food and pharmaceutical industries. The
citrus peel oils show how strong antimicrobial activity.

The antimicrobial activity has been checked in terms of

MIC by using different solvents against microorganism like
Pseudomonas aeuruginosa NCIM 2036 for which MIC was 1:20 in
presence of methanol, for Salmonella typhimurium NCIM 5021
the observed MIC was 1:20 in the presence of acetone. In
case of Micrococcus aureus NCIM 5021 the observed MIC was
1:20 when ethanol was used as solvent. The compounds like
coumarin and tetrazene were identified by GC/MS of lemon
peel extract. (Dhanavade, Jakulte, Sonawane, Ghosh,2011)

In the study entitled “Antimicrobial Activity of Aloe

Vera Leaf Extract” conducted by Kedarnath, Dr. Kaveri
Mahadevappa Kamble, Vishwanath B Chimkod, C S Pati. The
antimicrobial activity of aloe vera extract was tested
against pathogenic bacteria like Staphylococcus aureus,
Klebsiella pneumoniae and E.Coli and fungi like Aspergillus
niger and Candida at a dose of 1:20 mg/ml and 2:40 mg/ml by
using cup plate diffusion method. Various solvents such as
petroleum either, chloroform and methanol were used for
extracts. The results reveal that, methanol and petroleum
ether at a dose of 20mg/ml has showed significant activity
against Klebsiela pneumoniae and E.Coli whereas in fungi,
methanol extract showed significant activity against
Aspergillus niger and Candida.

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Methanol extract has showed maximum inhibitory activity

against E.Coli and Candida. Petroleum ether has showed
moderate inhibitory activity against Klebsiela pneumoniae
and Candida. The zone of inhibition was measured and
compared with standard Gentamycin (1mg/ml). However, in none
of the above mentioned extracts the inhibition zone was not
more than that found in standard i.e., Gentamycin.
(Kedarnath, Dr.Kamble, Chimkod, Patil, 2013)

CHAPTER III
MATERIALS AND METHODS

This chapter provides more intricate and important

details about the experiment. This also includes the

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materials needed and the procedures on how the experiment

will be conducted.
I. Research Design (Antibacterial activity)

This study will use the experiment type of research in

an Experimental Block Design. The study will utilize four
(4) treatments specifically treatment 1 as Aloe Vera gel
(Calamansi Zest + Virgin Coconut oil), treatment 2 as Aloe
Vera gel (Lemon + Virgin Coconut oil), treatment 3 as the
positive control (Green Cross Liquigel) and treatment 4 as
the negative control (distilled water). Each of the
treatments will test using Staphylococcus Aureus as the
bacterium. The antibacterial activity will identify by
measuring zone of inhibition of the four (4) treatments
using three (3) replicates.

Table 1. Treatments for Staphylococcus Aureus

Treatments Staphylococcus Aureus
Treatment 1: Aloe Vera Gel
(Calamansi Zest +
Virgin Coconut oil)

a. 25 % 1,2,3
b. 50 % 1,2,3
c. 75 % 1,2,3
Treatment 2: Aloe Vera Gel (Lemon
+ Virgin Coconut oil)
a. 25 % 1,2,3

b. 50% 1,2,3

c. 75 % 1,2,3

Treatment 3: Positive Control

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(Green Cross
Liquigel)

a. 25 % 1,2,3

b. 50 % 1,2,3

c. 75 % 1,2,3

Treatment 4: Negative Control

(Distilled water)

a. 25 % 1,2,3

b. 50 % 1,2,3

c. 75 % 1,2,3

II. Research Design (Microbial Analysis)

One hundred (100) grams of Aloe Vera Gel will present

at the Department of Science and Technology, Bagay Rd.,
Nursery Compound, Tuguegarao City, Cagayan for Mold Count
and Total Coliform Count.

NOTICE: There are different numbers of replicates in each

data of Mold count depending on what specific count you are
going to get and 1,2,3 is the standardized number in
identifying Mold Count of a given liquid gel.

Table 2. Treatments of ACLEO Liquigel for Mold Count and

Coliform count
Treatments Molds Count (petri- Coliform Count
plates) (tubes)

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Aloe Vera Gel

(Calamansi Zest +
Virgin Coconut oil)
a. 25 % 1,2,3,4 1,2,3

b. 50 % 1,2,3,4 1,2,3
c. 75 % 1,2,3,4 1,2,3

Aloe Vera Gel

(Lemon + Virgin
Coconut oil)
a. 25 % 1,2,3,4 1,2,3
b. 50 % 1,2,3,4 1,2,3
c. 75 % 1,2,3,4 1,2,3

III. SUBJECTS OF THE STUDY

-Staphylococcus Aureus bacterium will be used as the

subject of the study. Molds and Coliform are also the
subjects of this study that will be going to test from the
Department of Science and Technology, Bagay Rd., Nursery
Compound, Tuguegarao City, Cagayan, which there are
different numbers of replicates in each data depending on
what specific count you will be going to get and 1,2,3,4 is
the standardized number in identifying Mold Count of ACLEO
Liquigel.

IV. MATERIALS OF THE STUDY

This study will make possible due to the availability
of raw materials within and outside the locality. The
materials needed also in the preparation of ACLEO Liquigel
products can be found in every house’s kitchen.

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V. Preparation of ACLEO Liquigel Products

ACLEO Liquigel products were cooked, pasteurized, and
packaged in the Lemu National High School Gymnassium.
Sealing of the ACLEO Liquigel was made at Khenna Mineral
Water Station located in Divisoria, Enrile, Cagayan.

MATERIALS needed for ACLEO LIQUIGEL (aloe vera, calamansi

zest, lemon essential oil)

 100 grams of aloe vera  Clear glasses

 50 ml of calamansi zest  Small bowls
 5 ml of lemon essential  Measuring cups
oil  Weighing scale
 Knife  Dropper
 Chopping board  Beaker
 Clear container  Sealer
 Jelly powder

VI. Procedures

A. Aloe Vera Gel Extract

1. Fist prepare all the materials needed.
2. Do the antiseptic technique and wear the laboratory gears
such as gloves, facemask, and laboratory gowns.
3. Sterilize all the materials 10 to 15 minutes
4. Prepare the 5 pieces of aloe vera leaves
5. Clean off the cut aloe vera with clean water.
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6. Cut off the top third of the leaves.

7. Remove the spines also remove the top and bottom layers.
With the leaf lying flat on a cutting board, carefully slice
away the skin of the aloe vera leaf. This will be the thin,
outer green layer you see. Run your knife between it and the
thick, clear gel at the center, and repeat this process for
the remaining top layer by flipping it to the bottom.
8. Remove and store the gel. At this point, you should have
slabs of clear aloe vera gel. Be sure to rinse the aloe vera
gel itself 2 to 3 times when you’re finished to be sure that
it’s free of any aloe latex residue.
9. Store all the gel extract you get in a new glass or bowl
separate from the water you used to wash the leaves with.

B. Procedure in making Calamansi Zest

1. First prepare all the materials needed.
2. Sterilized the materials 10 to 15 minutes.
3. Prepare the calamansi fruit and wash it with clean water.
4. Use a grater with small holes and grate the calamansi
peeling over a bowl to get the calamansi zest.
5. After getting the calamansi zest place it in a clean
container.
C. Procedure in making Lemon Essential Oil
1. First prepare all the materials needed.
2. Sterilized the materials 10 to 15 minutes.
3. Prepare 2 to 3 lemons to be used.
4. Wash and dry the lemons. Rinse the lemons, scrub them
with a sponge or vegetable brush to remove pesticides and
dirt. Afterward, dry the lemons with a rag or paper towel.
5. Grate the skin of the lemon to get the lemon zest.

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6. Put the grated lemon in a small bowl and pour the virgin
coconut oil in the bowl. Make sure the oil will cover enough
the lemon zest.
7. Put water in a pot on the stove. Allow to heat up and
place your lemon zest with oil mixture and cooked it in a
low fire for about an hour.
8. Allow to cool and pour in a container. Leave in the
container to ferment for like one (1) week to get a better
result.
8. After a week pour the liquid through a strainer into a
bowl. The liquid in the bowl is your new lemon oil.
9. Store your lemon oil in an airtight container at room
temperature.

D. Mixing the Aloe Vera Gel, Calamansi Zest and Lemon

Essential Oil
1. Fist get the 100 grams of aloe vera in the bottle
container.
2. Put the 50 ml of calamansi zest to the aloe vera gel.
3. Mix the calamansi zest well with the aloe vera gel using
a small bowl with up to 5 minutes.
4. Get the 5 ml of lemon essential oil in the container.
5. Pour the 5 ml of lemon essential oil to the aloe vera gel
with calamansi zest and mix it well.
6. Sterilized the bottle container for 10 minutes.
7. Place the ACLEO Liquigel product in the bottle container.
8. Do the sealing process to avoid contamination of other
particulates.

FOR CLEANUP: Get all the materials that was used and place

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it on a medium-low heat of boil water. After which, throw

the mess and continue cleaning the other material with a
soap.

Proper waste disposal is needed by burying the residues on

the ground.

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